Compositions, methods and kits for treating a contact lens

ABSTRACT

Provided herein are fluorinated compounds having chemical structures: (I) where n is 0 or greater, or (II) where m and n are 0 or greater, or an amino acid, a soluble polymer, an oligo(ethylene glycol), a poly(ethylene glycol), or a carbohydrate each fluorinated with perfluorocarbons having the chemical structure (III) where n is 0 or greater. These fluorinated compounds are utilized in contact lenses to impart lipid-resistant, protein-resistant and biofouling-resistant properties, thus reducing discomfort and infection caused by contact lens wear without changing its transmission characteristics. Also provided is an ophthalmic drug delivery system comprising at least one of the compounds described above embedded in the contact lens and a kit to incorporate the ophthalmic into a contact lens. Methods for incorporating these compounds onto a contact lens without affecting transparency and for use in treating an ophthalmologic-associated condition are provided.

CROSS-REFERENCE TO RELATED APPLICATION

This application is a national stage application under 35 U.S.C. § 371of pending international application PCT/US2017/030065, filed Apr. 28,2017, which claims priority under 35 U.S.C. § 119(e) of provisionalapplication U.S. Ser. No. 62/329,388, filed Apr. 29, 2016, the entiretyof both of which are hereby incorporated by reference.

BACKGROUND OF THE INVENTION Field of the Invention

The present invention relates generally to the field of contact lenses.More specifically, the present invention relates to methods formodifying contact lenses into a drug delivery system. The presentinvention further relates to contact lenses which are lipid-resistant,protein-resistant and biofouling-resistant, thus reducing discomfort andinfection caused by contact lens wear.

Description of the Related Art

Currently more than 90% of ophthalmic drugs are delivered in the form ofeye drop solutions. Only ˜1-7% of the administered dose, however, isactively absorbed from such eye drop solutions. Therefore, high drugdosage and frequent administration are necessary in order to have atherapeutic effect, which results in undesired toxicity and low patientcompliance. Patient compliance is necessary to achieve desired treatmentefficacy, management of postoperative ocular pain and inflammation, andprevent chronic diseases such as glaucoma.

In the search for alternative approaches, contact lenses as ocular drugdelivery systems have attracted tremendous attention due to high oculardrug availability, less frequent administration, and low drug toxicity,which could potentially provide a more convenient treatment regimen andbetter patient compliance.

Approaches to incorporate drugs into contact lenses have evolved inrecent years, including simple immersion, covalent binding, molecularimprinting, layer-by-layer technique and incorporation with nanoscaledmaterials. Each method has its own advantages in one or two areas, butunfortunately, none to date have provided a simple manufacture process,high drug loading efficacy, satisfactory drug release kinetics, and highlens quality after drug incorporation despite rapid developments in thisfield.

Contact lenses have garnered significant popularity in the last fewdecades. Besides functional or optical reasons, many people choose towear contact lenses for aesthetic and cosmetic factors. However,discomfort and infection remains two major complications associated withcontact lens wear. Of the approximately 140 million contact lens wearersworldwide, 21-50% suffer from contact lens discomfort. This is the majorreason why individuals stop wearing contact lenses and is a majorchallenge to the contact lens industry.

Research has revealed that contact lens discomfort involves a series ofcomplex processes including, but not limited to, lipid deposition,oxidized lipid product deposition, protein deposition, proteindenaturation. These processes trigger host responses such as kinininflux and activation, and ocular mucin profile alternations. Theseprocesses are most likely caused by the disturbance of the nativeenvironment of ocular surface induced by the insertion of lens that isusually about ten times thicker than tear film.

Additionally, the absorption of tear film components onto the contactlens and/or the release of molecules from the contact lens to the tearfilm disturb the delicate balance of molecules in the tear film,resulting in tear film instability following a series of cellularresponses at the ocular surface. Further, contact lenses can serve as anincubator for microorganisms such as bacteria that increases the risk ofovert infection. Eye infection, which is often caused by adhesion andcolonization of bacteria such as Pseudomonas aeruginosa andStaphylococcus aureus, on contact lenses can be dangerous for a user'seyesight.

To reduce, or eliminate if possible, contact lens discomfort and contactlens infection, it is highly desirable to maintain the nativeenvironment of the ocular surface including the composition of lipids,proteins, and other components in the tear film, as well as theirbioactivity, even though the mechanical effects of the contact lens oneyelids and ocular surface are almost inevitable.

Silicone hydrogel lenses, one of the most commonly used types of contactlenses, are generally more hydrophobic, and thus attract more lipids andless proteins compared to other types of hydrogel lenses. However,proteins on silicone hydrogel lenses tend to denature more easily thanon other types of contact lenses, making protein an equal contributor aslipid to the discomfort when wearing silicone hydrogel contact lenses.

Extensive efforts have been made to reduce the discomfort and infectionsassociated with contact lens wear by limiting the deposition of lipid,protein and bacteria, including surface modification, plasma treatment,and optimization of lens material formulation, etc. To date, however,none of the methods are able to prevent all three types of depositionson contact lens due to different properties between these biomoleculesand/or bacteria that require drastically different treatment to resisttheir deposition on the contact lenses. For example, hydrophobicsurfaces may resist the adsorption of proteins but attract lipids.Another issue for current contact lens surface modification is polymerchain rearrangement, a process that changes surface properties ofcontact lenses over time, thus contributing to the failure of contactlenses to prevent the adsorption of biomolecules.

There is a recognized need, therefore, for a novel design of a contactlens drug delivery system and/or for methods for contact lens treatmentthat is able to resist the deposition of lipids, proteins and bacteriaand avoid the rearrangement of the polymer chains on the contactsurface. The present invention fulfills this longstanding need anddesire in the art.

SUMMARY OF THE INVENTION

The present invention is directed to a fluorinated compound effective asan ophthalmic drug and/or to reduce deposition of lipids, proteins andbacteria on a contact lens. The compound has the chemical structure:

wherein n is 0 or greater, or the chemical structure

wherein m and n are 0 or greater. The present invention is also directedto an amino acid, a soluble polymer, an oligo(ethylene glycol), apoly(ethylene glycol), or a carbohydrate each fluorinated with aperfluorocarbon having the chemical structure:

wherein n is 0 or greater.

The present invention also is directed to an ophthalmic drug deliverysystem comprising a contact lens embedded with at least one of thefluorinated compounds having structures described herein. The presentinvention is directed to a related ophthalmic drug delivery systemfurther comprising a saline solution rinse.

The present invention is directed further still to a method fordelivering an ophthalmic drug to a subject for treating anophthalmologic associated condition. The method comprises placing thecontact lens of the ophthalmic drug delivery system described hereinonto an eye of the subject. The at least one fluorinated compounds aredelivered to the eye thereby treating the opthalmologic-associatedcondition. The present invention is directed to a related method furthercomprising rinsing the contact lens prior to placing it onto the eye.

The present invention is directed further still to a kit to reducedeposition of lipids, proteins and bacteria on a contact lens. The kitcomprises a solution of the fluorinated compounds as described hereinand instructions to use the kit. The present invention is directed to arelated kit further comprising a saline solution rinse. The presentinvention is directed to another related kit further comprising asterile and sealable vial for holding the solution of fluorinatedcompound(s).

The present invention is directed further still to a method formodifying a contact lens to deliver drugs and/or to reduce deposition oflipids, proteins, and bacteria on a contact lens. The method comprisesthe steps of drying the contact lens, incubating the lens with one ormore fluorinated compounds in a medium and rinsing the lens.

The present invention is directed further still to a modified contactlens produced by the method described herein.

The present invention is directed further still to a method forincorporating ophthalmic drugs and/or for reducing the deposition oflipids, proteins, and bacteria on a contact lens. This method comprisesthe steps of producing the contact lens, incubating the contact lens ina solution of a fluorinated zwitterionic compound and/or a fluorinatedophthalmic drug compound and rinsing the contact lens. The presentinvention is directed to a related method further comprising storing thecontact lens.

The present invention is directed further still to a method formaintaining transparency in a contact lens. The method comprises thesteps of applying a fluorinated zwitterionic compound and/or afluorinated antibacterial compound in a medium to the contact lens andrinsing the contact lens.

Other and further aspects, features, and advantages of the presentinvention will be apparent from the following description of thepresently preferred embodiments of the invention given for the purposeof disclosure.

BRIEF DESCRIPTION OF THE DRAWINGS

So that the matter in which the above-recited features, advantages andobjects of the invention, as well as others that will become clear, areattained and can be understood in detail, more particular descriptionsof the invention briefly summarized above may be had by reference tocertain embodiments thereof that are illustrated in the appendeddrawings. These drawings form a part of the specification. It is to benoted, however, that the appended drawings illustrate preferredembodiments of the invention and therefore are not to be consideredlimiting in their scope.

FIG. 1 shows the synthesis scheme for fluorinated Compound 1. Compounds3 and 4 are used to produce compound 5. Then compounds 5 and 2 arecombined at the molar ratio of 1:1 produce the fluorinated zwitterioniccompound 1.

FIG. 2 shows the chemical structures of molecules tested to attach tocontact lenses. A fluorescent dye tagged with a fluorinated carbon chain(FITC-C₈F_(n)) shown in FIG. 2 is used as a model compound 6 (n=5). Theattachment of these compounds to the contact lenses can be easilydetected and quantified by its fluorescence signal. Compound 7 (FITC) isused as negative control. FIG. 2 . also shows the structure offluorinated ciprofloxacin compounds 8-11 (F-Cip 1-4) and compound 12(Cip).

FIG. 3 illustrates the amount of compound 6 (FITC-C₈F₁₅) and compound 7(FITC) loaded onto Comfilcon A, Lotrafilcon B, Delefilcon A, NarafilconA and Ocufilcon D lenses. Data are from at least three independentexperiments and are expressed as mean±SD. Two-way ANOVA is performedfollowed by Tukey's test where significance is found. *p<0.05.

FIG. 4A-4B Illustrate contact lens transparency after incubation withF-Cip compounds. FIG. 4A shows Comfilcon A lens transmission between 400nm and 700 nm after loading with compound 8 (F-Cip1) and compound 12(Cip). FIG. 4B lens transparency within a wavelength range of 220-800 nmin Comfilcon A lenses loaded with F-Cip1, F-Cip2, Cip. Transmissionthrough unloaded fresh lens is shown for comparison.

FIG. 5A-5E shows release of compounds from Comfilcon A lens. FIG. 5Aillustrates the amount of compound 6 released over 16 h at a solutionexchange rate of 1 mL/h. FIG. 5B illustrates the amount of compound 7released over 8 h at a solution exchange rate of 1 mL/h. FIG. 5Cillustrates the amount of compound 6 released over 360 h at a solutionexchange rate of 6 mL/h. FIG. 5D illustrates the amount of compound 7released over 120 h at a solution exchange rate of 6 mL/h. Data areexpressed as mean±SD from three or more independent experiments. FIG. 5Eshows release of F-Cip1 (compound 8), F-Cip2 (compound 9) and Cip(compound 12) from Comfilcon A lenses over a course of 8 h at a solutionexchange rate of 1 mL/h. Data are expressed as mean±SD from three ormore independent experiments.

FIG. 6 illustrates the amount of F-Cip compounds 8, 9, 10 and 11, andCip, (compound 12) loaded onto Comfilcon A lenses. Data are mean±SD fromthree independent experiments. One-way ANOVA was performed followed byTukey's test where significance was found. *p<0.05.

FIG. 7 shows antimicrobial activity against P. aeruginosa 19660 inComfilcon A lenses loaded with compounds 8 and 9 and control compound12. Data are from three independent experiments expressed as mean±SD.One-way ANOVA is performed followed by Tukey's test where significanceis found *p<0.05.

FIG. 8 shows antimicrobial efficacy of modified Comfilcon A lenses in anex vivo porcine eye infection model. CL: unmodified lenses. 1: lensesmodified with F-Cip 1. Cip: lenses modified with Cip. 0.3% Cip: 0.3%ciprofloxacin solution administered in the form of eye drops. Noscratch: intact porcine eyes. Statistical analysis is performed usingone-way ANOVA followed by Tukey's test comparing to 1 where significanceis found. *p<0.05. (n. 3).

FIGS. 9A-9B illustrate the cytotoxicity of compounds. FIG. 9A showscytotoxicity of compounds 8 and 9 and compound 12 (control) againsthuman telomerase corneal epithelial cells (hTCEpi) at a series ofconcentrations. Benzalkonium chloride (BAC) is used as positive control.Data are expressed as mean±SD from three independent experiments. FIG.9B shows cytotoxicity of compounds 8 and 9 and compound 12(control)-loaded Comfilcon A lenses against human telomerase cornealepithelial cells (hTCEpi). Data are mean±SD from three independentexperiments.

DETAILED DESCRIPTION OF THE INVENTION

As used herein in the specification, “a” or “an” may mean one or more.As used herein in the claim(s), when used in conjunction with the word“comprising”, the words “a” or “an” may mean one or more than one.

As used herein “another” or “other” may mean at least a second or moreof the same or different claim element or components thereof. Similarly,the word “or” is intended to include “and” unless the context clearlyindicates otherwise. “Comprise” means “include.”

As used herein, the term “about” refers to a numeric value, including,for example, whole numbers, fractions, and percentages, whether or notexplicitly indicated. The term “about” generally refers to a range ofnumerical values (e.g., +/−5-10% of the recited value) that one ofordinary skill in the art would consider equivalent to the recited value(e.g., having the same function or result). In some instances, the term“about” may include numerical values that are rounded to the nearestsignificant figure.

As used herein, the term “blister pack” refers to a plastic, bubble-likepocket to store contact lens. Blister packs typically have a liddingseal of aluminum foil.

In one embodiment of the present invention there is provided afluorinated compound effective as an ophthalmic drug and/or to reducedeposition of lipids, proteins and bacteria on a contact lens having thechemical structure:

where n is 0 or greater; or

wherein n is 0 or greater and m is 0 or greater. Representative examplesof useful fluorinated compounds include but are not limited to compound8, compound 9, compound 10 or compound 11.

Alternative compounds are amino acids, a soluble polymer, anoligo(ethylene glycol), a poly(ethylene glycol), or a carbohydrate eachfluorinated with a perfluorocarbon having the chemical structure:

where n is 0 or greater. These compounds may be incorporated singly orin combination with each other and/or with the described fluorinatedzwitterionic compounds.

The fluorinated chain lengths may be increased and the surface densityof the zwitterionic groups may be adjusted. This can maximize theresistance of the contact lens to lipids, proteins, and microorganismsand the time span for which the molecules are in contact with thelenses. As the tear film is a complex system of lipids, proteins,enzymes, etc., the long-term interactions between the modified contactlenses and the tear film could possibly detach those molecules attachedto the contact lenses, leading to loss of resistance of the contact lensto the tear film components (lipids, proteins) as well as microorganismsthereby making these contact lenses comfortable to wear. Optimizing thechain length can strengthen the interaction between the fluorinatedzwitterionic molecules and the contact lenses, resulting in highersurface density and a longer time span that the molecules stay attachedon the contact lenses.

In another embodiment of the present invention there is provided anophthalmic drug delivery system comprising a contact lens embedded withat least one of the fluorinated compounds having structures shown above.Further to this embodiment the ophthalmic drug delivery system furthercomprises a saline solution rinse.

In yet another embodiment of the present invention, there is provided amethod for delivering an ophthalmic drug to a subject for treating anopthalmologic-associated condition, comprising placing the contact lensof the ophthalmic drug delivery system as described supra onto an eye ofthe subject, where the at least one fluorinated compounds are deliveredto the eye thereby treating the opthalmologic-associated condition. In afurther embodiment the method comprises rinsing the contact lens priorto the placing step. In both embodiments the opthalmologic-associatedcondition may be an infection, glaucoma, inflammation, an allergy, orpain.

In yet another embodiment of the present invention there is provided akit to incorporate ophthalmic drugs and/or to reduce deposition oflipids, proteins and bacteria on a contact lens, comprising a solutionof the fluorinated zwitterionic compound or fluorinated ophthalmic drugsas described supra or an amino acid, a soluble polymer, an oligo(ethylene glycol), a poly(ethylene glycol), or a carbohydrate eachfluorinated with a perfluorocarbon having the chemical structure:

where n is 0 or greater or a combination thereof; and instructions touse the kit.

Further to this embodiment, the kit comprises a saline solution rinse.In another further embodiment, the kit comprises a sterile and sealablevial for holding the solution. In all embodiments, the solution maycomprise a saline or an organic solvent. Preferably, the saline isphosphate buffered saline, saline, contact lens storage solution, andcontact lens cleaning solution and the organic solvent istetrahydrofuran, ethanol, or isopropanol.

In another embodiment of the present invention, there is provided amethod for modifying a contact lens to deliver drugs and/or to reducethe deposition of lipids, proteins, and bacteria on a contact lens thatcontains fluorine, comprising the steps of drying the contact lens;incubating the contact lens with a medium containing a one or morefluorinated compounds; and rinsing the contact lens. In this embodiment,a representative incubating medium is a buffer. Representative buffersinclude, but are not limited to, phosphate buffered saline, saline,contact lens storage solution, and contact lens cleaning solution.Alternatively, a useful medium is an organic solvent. Representativeorganic solvents include, but are not limited to, tetrahydrofuran,ethanol, and isopropanol.

Generally, the contact lens is incubated for an amount of time necessaryto incorporate the fluorinated compound(s) for drug delivery and/orreducing the deposition of lipids, proteins, and bacteria on a contactlens. Preferably, the contact lens is incubated for about 4 hours toabout 24 hours, although a person having ordinary skill in this artwould readily recognize that the incubation time could be as short asseconds, or as long as its entire shelf life, depending on the methodutilized to do the job. The incubation is preferably performed at roomtemperature. In this embodiment, the method can prevent the polymerchain rearrangement of the contact lens and further, maintaintransparency of the contact lens, provide ophthalmic drugs for deliveryand make the contact lens resistant to microbial agents afterincubation.

Particularly, the fluorinated compound is a fluorinated zwitterioniccompound having the chemical structure

where n is 0 or greater, a fluorinated ophthalmic drug compound havingthe chemical structure

wherein n is 0 or greater and m is 0 or greater; or other fluorinatedophthalmic drugs, an amino acid, a soluble polymer, an oligo(ethyleneglycol), a poly(ethylene glycol), or a carbohydrate each fluorinatedwith a perfluorocarbon having the chemical structure:

wherein n is 0 or greater. A representative fluorinated ciprofloxacincompound may be compound 8 or compound 9. For example, the fluorinatedciprofloxacin has a value of 2 for n and a value of 0 for m and has thestructure of compound 8 shown in FIG. 2 , wherein incubating the contactlens with a solution containing this compound makes the lens resistantto microbial agents. Alternatively, the fluorinated ciprofloxacin has avalue of 3 for n and a value of 0 for m and has the structure ofcompound 9 shown FIG. 2 , wherein incubating the contact lens with asolution containing this compound makes the lens resistant to microbialagents. Alternatively, the fluorinated ciprofloxacin has a value of 5for n and a value of 0 for m with structure of compound 10 shown FIG. 2, or, a value of 1 for n and a value of 2 for m having the structure ofcompound 11 shown FIG. 2 , wherein incubating the contact lens with asolution containing either compound makes the lens resistant tomicrobial agents.

In a related embodiment of the present invention, there is provided amodified contact lens produced by the method as described supra.

In yet another embodiment of the present invention, there is provided amethod for incorporating ophthalmic drugs and/or for reducing thedeposition of lipids, proteins, and bacteria on a contact lens in amanufacturing process, comprising the steps of producing the contactlens; incubating the contact lens in a solution of a fluorinatedzwitterionic compound and/or a fluorinated ophthalmic drug compound; andrinsing the contact lens. Further to this embodiment the methodcomprises storing the contact lens.

In both embodiments the fluorinated compound may be a fluorinatedzwitterionic compound having the chemical structure

wherein n is 0 or greater. Alternatively, in both embodiments thefluorinated ophthalmic drug compound has the chemical structure:

wherein n is 0 or greater and m is 0 or greater as described supra.

In both embodiments, the solution may comprise phosphate buffered salineor tetrahydrofuran, ethanol or isopropanol. In addition, the contactlens may be incubated about 4 hours to about 24 hours.

In yet another embodiment of the present invention, there is provided amethod for maintaining transparency in a contact lens, comprising thesteps of applying a fluorinated zwitterionic compound and/or afluorinated antibacterial compound in a medium to the contact lens; andrinsing the contact lens. In this embodiment the fluorinatedzwitterionic compound or a fluorinated antibacterial compound, forexample, ciprofloxacin are as described supra. In this embodiment themedium may comprise phosphate buffered saline or tetrahydrofuran,ethanol or isopropanol. Typically, the fluorinated compounds may beapplied to the contact lens for about 4 hours to about 24 hours.

In yet another embodiment of the present invention, there is provided amethod for increasing a level of comfort and safety of a contact lensfor a user thereof, comprising the steps of incubating the contact lensin a medium comprising a fluorinated zwitterionic compound and/or afluorinated antibacterial compound to reduce the deposition of lipids,proteins, and bacteria thereon; and rinsing the contact lens. In thisembodiment the fluorinated zwitterionic compound or the fluorinatedantibacterial compound and the medium are as described supra. Also, thecontact lens may be rinsed after about 4 hours to about 24 hours.

Provided herein are methods, fluorinated compounds, includingfluorinated zwitterionic compounds and fluorinated ophthalmic drugcompounds, and kits comprising the same for incorporating the ophthalmicdrugs into contact lenses and/or for increasing the resistance tolipids, proteins, and bacteria on the surface of contact lenses. Theincreased resistance of the contact lens prevents loss of transparencyin a contact lens, increases a level of comfort and safety of a contactlens for a user or wearer thereof and increases resistance to microbialor bacterial activity.

These methods can be used for any contact lens containing fluorine, suchas, but not limited to Comfilcon A and Enfilcon A contact lenses.Particularly, the fluorinated compounds are fluorinated zwitterioniccompounds such as shown synthesized using the scheme in FIG. 1 or afluorinated anti-bacterial or anti-microbial agent, such as, fluorinatedciprofloxacin derivatives as shown in FIG. 2 , or other fluorinatedophthalmic drug compound. The methods and fluorinated compounds providedherein are used as a final step of an industrial manufacturing processfor contact lens production or as a post-production step initiated by auser or wearer of the contact lens. In a packing procedure the medium orsolution comprising the fluorinated compounds may be stored in a blisterpack for example or contained as an additive in the traditional contactlens cleaning solution for daily usage. Thus, also provided is amodified contact lens produced to contain or incorporate one or more ofthe described fluorinated compounds.

Particularly, the zwitterionic compounds attach to the fluorinatedmolecules in the contact lenses via a strong and specific fluorousattraction. The fluorinated compounds are neither water-friendly noroil-friendly. A layer applied, coated or disposed on the outermost layerof the contact lens via fluorous attraction prevents the lipid andprotein deposition. Also, when attached to the contact lenses, thezwitterionic heads of the compounds on the outer layer of the contactlenses and the fluorinated chains are embedded in the contact lens. Thepolymer chain rearrangement may be minimized since the fluorinatedcarbon chains are neither hydrophilic nor hydrophobic. The zwitterionicheads of the compounds prevent the deposition of microorganisms.

Alternative compounds are amino acids, a soluble polymer, anoligo(ethylene glycol), a poly(ethylene glycol), or a carbohydrate eachfluorinated with a perfluorocarbon having the chemical structure:

wherein n is 0 or greater. These compounds may be incorporated singly orin combination with each other and/or with the described fluorinatedzwitterionic compounds.

The fluorinated chain lengths may be increased and the surface densityof the zwitterionic groups may be adjusted. This can maximize theresistance of the contact lens to lipids, proteins, and microorganismsand the time span for which the molecules are in contact with thelenses. As the tear film is a complex system of lipids, proteins,enzymes, etc., the long-term interactions between the modified contactlenses and the tear film could possibly detach those molecules attachedto the contact lenses, leading to loss of resistance of the contact lensto the tear film components (lipids, proteins) as well as microorganismsthereby making these contact lenses comfortable to wear. Optimizing thechain length can strengthen the interaction between the fluorinatedzwitterionic molecules and the contact lenses, resulting in highersurface density and a longer time span that the molecules stay attachedon the contact lenses.

A representative incubating medium is a buffer. Representative buffersinclude, but are not limited to, phosphate buffered saline, saline,contact lens storage solution, and contact lens cleaning solution.Alternatively, a useful medium is an organic solvent. Representativeorganic solvents include, but are not limited to, tetrahydrofuran,ethanol, and isopropanol. Tetrahydrofuran, may be used to replacephosphate buffered saline to dissolve the fluorinated zwitterioniccompound since the solubility of the fluorinated zwitterionic compoundmay become limited in phosphate buffered saline depending on the lengthof fluorinated carbon chain.

The fluorinated compounds also may comprise a fluorinated antibacterial,anti-microbial or ophthalmic drug compounds which when incorporated intothe contact lens via incubation or other application produces a drugdelivery system in a modified contact lens that is worn by a subject oruser thereof. The modified contact lens is useful to treat anophthalmic-associated condition, disease or disorder. These therapeuticsmay be incorporated with or without the fluorinated zwitterioniccompounds. The therapeutics in the modified contact lens are eluted viathe natural tearing action of the eye. These therapeutic ophthalmiccompounds can be used to treat and/or alleviate conditions such asinfection, glaucoma, inflammation, allergy, or pain associated therewithwhen the contact lens comprising the one or more ophthalmic therapeuticsis worn in one or both eyes.

Contact lens care solutions containing the above fluorinated drug andlubricating molecules can be used for storage of commercial fluorinatedcontact lenses between wear. Due to the fluorous interactions, thesemolecules will incorporate into contact lenses and provide therapeuticor improved wear experience.

The invention thus provides a number of advantages and uses, which aredescribed below, however, the advantages and uses are not limited bythis description. Embodiments of the present invention are betterillustrated with reference to the Figure(s), however, such reference isnot meant to limit the present invention in any fashion. The embodimentsand variations described in detail herein are to be interpreted by theappended claims and equivalents thereof.

Example 1

Synthesis of Fluorinated Molecules

The synthetic scheme for fluorinated molecules 1 is shown in FIG. 1 .For compound 2 the n in the formulation is equal to 7. Compounds 2 and 3are used without further purification. Compounds 3 and 4 with a molarratio of 1:1.1 are dissolved in tetrahydrofuran in a round shape flaskand stirred using a magnetic stirrer at room temperature overnight. Thesolvent is evaporated using a rotary evaporator, and the product 5 ispurified using chromatography. A mixture of 2 and 5 with a molar ratioof 1:1 is stirred at room temperature overnight followed by solventremoval and purification using chromatography. All purified compoundsare characterized using mass spectroscopy and nuclear magnetic resonance(NMR).

Synthesis of Fluorinated Fluorescein

The synthesis of FITC-C₈F₁₅ is carried out as reported previously.Briefly, 15.57 mg of fluorescein isothiocyanate (FITC) is dissolved in10 mL of ethanol. Then, 10 mL of 1H, 1H-perfluorooctylamine (2 mM inethanol) is added drop wise at room temperature with stirring. Thesolution is stirred at room temperature for 24 hours. The solvent isthen evaporated under reduced pressure, and the residue purified bysilica gel flash chromatography (ethyl acetate/Methanol 9/1), to giveFITC—C₈F₁₅ (22 mg, 93% yield) as a green powder (FIG. 2 ). The structureof FITC-C₈F₁₅ is verified by NMR and MALDI-TOF-MS m/z: [M]⁺ to have theformula, C₂₉H₁₇F₁₅N₂O₅S, and a formula weight of 790.06219.

¹H NMR (500 MHz, acetone-d6) δ 9.05 (s, 2H), 7.98-7.89 (m, 2H), 7.26(dd, J=8.1, 7.0 Hz, 1 H), 6.75 (d, J=2.5 Hz, 2H), 6.72 (d, J=4.0 Hz,1H), 6.70 (d, J=3.5 Hz, 1H), 6.65-6.64 (m, 1H), 6.63 (dt, J=4.9, 1.8 Hz,1H), 4.75 (dtd, J=48.0,16.4, 6.2 Hz, 2H), 4.47 (q, J=7.2 Hz, 1H). ¹³CNMR (126 MHz, Acetone-d6) δ 184.39, 169.11, 169.07, 160.30, 153.35,153.32, 153.28, 149.98, 146.63, 141.81, 131.30, 130.17, 130.13, 130.09,128.40, 125.88, 125.17, 125.12, 124.75, 119.22, 119.07, 117.36, 116.63,113.30, 111.65, 111.58, 111.55, 103.33, 55.44. ¹⁹F NMR (471 MHz, CDCl₃)δ −81.44-81.64 (m, 3F), −117.09-117.92 (m, 2F), −122.23 (s, 2F), −122.46(s, 2F), −123.18 (s, 2F), −123.94 (s, 2F), −126.63 (td, J=14.6, 6.9 Hz,2F). MALDI-TOF-MS m/z: [M]⁺ calculated for C₂₉H₁₇F₁₅N₂O₅S=790.06; found:790.10.

Synthesis of Fluorinated Ciprofloxacin Derivatives

Ciprofloxacin (250 mg, 0.75 mmol) and triethylamine (139 μL, 1 mmol) isstirred in anhydrous methylene chloride (5 mL) at 0° C. for 15 min.Heptafluorobutyric acyl chloride (259.8 mg, 1.12 mmol) is added dropwise to the mixture and the reaction is protected under nitrogenatmosphere. The suspension is stirred at room temperature for 12 hours,following which, the volatile components are removed under reducedpressure. The residue is then purified by silica gel columnchromatography to yield compound 8 (173.9 mg, 33% yield) as a whitepowder (FIG. 2 m=0, n=2) verified by NMR and MS (ESI) m/z: [M+H]⁺ tohave the formula C₂₁H₁₈F₈N₃O₄ and a formula weight of 528.09.

¹H NMR (500 MHz, CDCl₃) d 8.73 (s, 1H), 8.01 (d, J=12.7 Hz, 1H), 7.38(d, J=7.0 Hz, 1H), 3.96 (dd, J=8.9, 4.7 Hz, 4H), 3.56 (s, 1H), 3.45-3.36(m, 4H), 1.42 (q, J=6.6 Hz, 2H), 1.21 (t, J=5.0 Hz, 2H). ¹³C NMR (126MHz, CDCl₃) δ 177.15, 166.83, 156.47, 154.70, 152.70, 147.81, 145.03,139.05, 120.82, 112.95, 112.77, 108.40, 105.50, 50.17, 49.42, 45.95,43.55, 35.49, 8.43. ¹⁹F NMR (470 MHz, CDCl₃) δ 79.58 (t, J=9.5 Hz, 3F),−111.46-111.53 (m, 2F), −121.14-121.21 (m, 2F), −125.51-125.57 (m, 2F).MS (ESI) m/z: [M+H]⁺ calculated for C₂₁H₈F₈N₃O₄=528.12; found 528.09.

Ciprofloxacin (250 mg, 0.75 mmol) and triethylamine (139 μL, 1 mmol) arestirred in anhydrous methylene chloride (5 mL) at 0° C. for 15 min.Perfluoropentanoyl chloride (315.8 mg, 1.12 mmol) is added drop wiseinto the mixture and the reaction is protected under nitrogenatmosphere. The suspension is stirred at room temperature for 12 hours,following which, the volatile components are removed under reducedpressure and the residue further purified by silica gel columnchromatography to yield Compound 9 (178.9 mg, 31% yield) as a yellowpowder (FIG. 2 m=0, n=3) verified by NMR and MS (ESI) m/z: [M+H]⁺ tohave the formula of C₂₂H₁₈F₁₀N₃O₄=578.11 and a formula weight of 578.08.

¹H NMR (500 MHz, Acetone-d6) δ 8.70 (s, 1H), 7.93 (d, J=13.1 Hz, 1H),7.79 (d, J=7.4 Hz, 1H), 4.06-3.94 (m, 4H), 3.88 (dd, J=7.1, 3.5 Hz, 1H),3.61-3.52 (m, 4H), 1.48 (d, J=6.0 Hz, 2H), 1.39-1.30 (m, 2H). ¹³C NMR(126 MHz, Acetone-d6) δ 177.89, 166.64, 156.59, 155.42, 153.44, 149.00,145.87, 145.79, 140.35, 130.20, 129.37, 127.17, 120.96, 112.31, 112.12,108.59, 107.80, 50.73, 50.05, 46.41, 44.04, 36.61, 8.49. ¹⁹F NMR (471MHz, CDCI₃) δ 80.86 (t, J=9.8 Hz, 3F), δ −110.94 (t, J=12.2 Hz, 2F),_121.09 (dd, J=12.7, 6.9 Hz, 2F), _121.86 (dddd, J=13.4, 10.0, 6.7, 3.4Hz, 2F), 124.50-24.62 (m, 2F). MS (ESI) m/z: [M+H]⁺ calculated forC₂₂H₁₈F₁₀N₃O₄=578.11; found 578.08.

Ciprofloxacin (250 mg, 0.75 mmol) and triethylamine (139 μL, 1 mmol) arestirred in anhydrous methylene chloride (5 mL) at 0° C. for 15 min. Theperfluoroheptanoic acyl chloride (427.8 mg, 1.12 mmol) is added dropwise in to the mixture and the reaction is protected under nitrogenatmosphere. The suspension is stirred at room temperature for 12 hours,following which, the volatile components are removed under reducedpressure and the residue further purified by silica gel columnchromatography to yield compound 10 (202 mg, 40% yield) as a pale yellowpowder.

¹H NMR (500 MHz, CDCl₃) δ 8.63 (s, 1H), 7.86 (d, J=12.7 Hz, 1H), 7.34(d, J=7.0 Hz, 1H), 3.93-3.71 (m, 4H), 3.61-3.51 (m, 1H), 3.47-3.28 (m,4H), 2.68 (dd, J=9.6, 6.2 Hz, 2H), 2.49 (ddd, J=17.6, 14.4, 7.8 Hz, 2H),1.40 (q, J=6.7 Hz, 2H), 1.20 (q, J=6.5 Hz, 2H). ¹³C NMR (126 MHz, CDCl₃)δ 176.91, 168.77, 167.31, 154.64, 152.64, 147.63, 145.46, 145.37,139.06, 120.04, 119.98, 112.50, 112.31, 107.84, 105.24, 49.89, 49.36,45.23, 41.67, 35.53, 26.57, 26.40, 26.23, 24.38, 8.32. ¹⁹F NMR (470 MHz,CDC13) δ 80.62 (t, J=9.9 Hz, 3F), −110.68 (t, J=13.4 Hz, 2F), −120.60(s, 2F), −121.00 (dd, J=15.2, 8.0 Hz, 2F), −121.14 (dd, J=12.7, 6.9 Hz,2F), −122.67 (s, 2F), −125.81-125.92 (m, 2F). MS (ESI): [M+H]⁺calculated for C₂₂H₂₂F₆N₃O₄=506.1, Found 506.2.

Ciprofloxacin (250 mg, 0.75 mmol) and triethylamine (139 μL, 1 mmol) arestirred in anhydrous methylene chloride (5 mL) at 0° C. for 15 min. The4,4,5,5,5-pentafluoropentanoic acyl chloride (235.2 mg, 1.12 mmol) isadded drop wise in to the mixture and the reaction is protected undernitrogen atmosphere. The suspension is stirred at room temperature for12 hours, following which, the volatile components are removed underreduced pressure and the residue further purified by silica gel columnchromatography to yield compound 11 (203 mg, 30% yield) as a yellowpowder.

¹H NMR (500 MHz, CDCl₃) δ 8.77 (s, 1H), 8.05 (d, J=12.7 Hz, 1H), 7.38(d, J=6.6 Hz, 1H), 3.96 (d, J=11.8 Hz, 4H), 3.55 (s, 1H), 3.40 (s, 4H),1.42 (d, J=5.9 Hz, 2H), 1.22 (s, 2H). ¹³C NMR (126 MHz, CDCl₃) δ 177.23,166.85, 156.57, 154.73, 152.73, 147.87, 144.95, 139.08, 121.04, 120.98,113.10, 112.92, 111.37, 110.84, 110.62, 108.56, 105.50, 50.26, 49.45,46.00, 43.62, 35.48, 8.45. ¹⁹F NMR (470 MHz, CDC13) δ −85.32 (s, 3F),−118.30 (t, J=18.5 Hz, 2F), −121.05-121.10 (m, 2F). MS (ESI): [M+H]⁺calculated for C₂₄H₁₈F₁₄N₃O₄=678.1, Found 678.1.

Example 2

Modification of Contact Lens with Fluorinated Molecules

Commercial Comfilcon A lenses are removed from blister packs, andblotted gently to remove excess solution. Lenses are placed individuallyinto glass vials containing the synthesized fluorinated molecules inphosphate buffered saline or other saline (20 μM) and incubated for 4hours to 24 hours at room temperature. At the end of incubation, lensesare washed with PBS or saline three times, and blotted dry or air driedfor further experiments. As the fluorinated carbon chain lengthincreases, the solubility of these molecules will become limited. Ifnecessary, organic solvent is used for better solubility.

Example 3

Quantification of Attached Molecules and Characterization of theModified Contact Lens

To determine the amount of fluorinated molecules attached onto contactlenses, perfluorocarbon solvent is used to dissolve the attachedmolecules out, and liquid chromatography-Mass Spectrometry is used toidentify and quantify the amount. Specifically, modified lenses areplaced into glass vials containing 1 mL perfluorocarbon solvent andsonicated for several minutes at room temperature. This process isrepeated 3 times, and then all the solution is collected and combinedinto a flask followed by evaporation using a rotary evaporator. Theresidue in the flask is dissolved in 100 μL of acetonitrile and injectedinto liquid chromatography-Mass Spectrometry with acetonitrile as mobilephase. The molecule is identified from Mass Spectrometry spectrum usingmolecular ion peak and the quantity is measured from the chromatographypeak area.

X-ray photoelectron spectroscopy (XPS) is used to determine the surfaceproperties and atomic concentrations of C1s, N1s, O1s, F1s, Si2p of fivetypes of commercial contact lenses, Comfilcon A, Narafilcon A,Lotrafilcon B, Ocufilcon D, and Delefilcon A, and contact angle aremeasured by goniometer. Briefly, lenses are dried in air and cut intopieces of ˜5×5 mm². Each piece is glued onto a stainless steel holderand loaded into a PHI 5700 X-ray photoelectron spectrometer, equippedwith a monochromatic AlKα X-ray source (hv=1486.7 eV) at a take-offangle (TOA) of 45° from the film surface. XPS confirms the presence offluorinated molecules at the surface of contact lenses by detectingfluorine signals. The advancing and receding contact angles are measuredusing a Rame-Hart goniometer. At least four drops of probe liquid aremeasured for each sample.

Example 4

Statistical Analysis

Data are expressed as mean±SD. Where applicable, statistical analysis isperformed using one-way or two-way ANOVA followed by Tukey's test wheresignificance is found with p<0.05.

Adsorption of Lipids, Proteins and Microorganisms onto Modified ContactLenses

The adsorption of lipids such as phosphatidylcholine and cholesterololeate, proteins such as lysozyme and albumin, and microorganisms suchas Pseudomonas aeruginosa and Staphylococcus aureus are tested oncontact lenses modified above.

Loading of Molecules into Commercial Contact Lenses

A list of the commercial contact lenses tested is presented in Table 1.All lenses are air dried and weighed. The weights of Comfilcon A,Narafilcon A, Lotrafilcon B, Ocufilcon D and Delefilcon A lenses were16.3±0.3, 15.6±0.1, 20.7±0.3, 15.7±0.2, and 21.5±0.4 mg, respectively.Compounds used including FITC-F, FITC, Ciprofloxacin (Cip, compound 12),fluorous-tagged ciprofloxacin (F-Cip1-4) are shown in FIG. 2 .

Briefly, commercial lenses are individually immersed in wells of a 24well plate, each containing 1 mL PBS solution of FITC-F (15.6 mM), FITC(15.6 mM), Cip (100 mM) or F-Cip (100 mM), and incubated for 18 h. Thenthe lenses are washed with PBS three times for 5 min each. Allincubation and wash solutions are collected and the amount of eachcompound is quantified by spectrophotometry. Standard curves aregenerated with seven standard samples with known concentrations for eachindividual compound of interest. Specifically, fluorescent compoundsFITC and FITC-F are quantified by fluorescence emission at 520 nm(lex.485 nm), and Cip and F-Cip are quantified by absorbance at 275 nm.The amount of each compound in the above collected solutions is thenmeasured with the standard curve. The amount of the compoundincorporated into lenses was determined by subtracting the amount insolution from the total amount added initially.

Lens Transparency After Modification

To determine the transparency of fresh and modified Comfilcon A contactlenses using light transmission, contact lenses are cut into small diskswith a diameter of 6 mm and placed into wells of a UV-transparent96-well plate, and a wavelength scan from 220 to 800 nm is conductedusing a plate reader (BMG LabTech FLUOstar Omega plate reader, Germany).

Time Course Release of Individual Compounds from Contact Lenses

The modified Comfilcon A lenses are placed into individual wells of a 24well plate with 1 mL PBS in each well at room temperature. The PBSsolution is replaced with fresh PBS every hour for up to 8 h or 15 h, orevery 10 min for 2 h or 6 h. The amount of fluorescent compound releasedinto the collected PBS aliquots is quantified spectrophotometricallybased on the above standard curve. The amount of F-Cip released into thecollected PBS aliquots is quantified using LC-MS (LCQ Deca XP plus withSurveyor LC, Thermo Fisher, Waltham, Mass.) with electrospray ionizationin positive ion mode. Samples (5 mL each) are loaded onto the analyticalcolumn at 200 mL/min (Kinetex XB-C18, 2.×50 mm, 2.6 mm, Phenomenex) withthe following gradient: 0-3 min, 10-75% (B); 3-7.5 min, 75-91% (B). Themobile phase consists of (A) water containing 0.1% formic acid and (B)acetonitrile containing 0.1% formic acid. Mass spectrometric parametersare as follows: 250° C. capillary temperature, 45 units sheath gas, 10units aux gas and 4.5 kV spray voltage. Selective ion monitoring (SIM)is used for detecting F-Cip 1 and 2 (compounds 8 and 9), and thequantification is based on the respective standard calibration curve.The detection limit for F-Cip 1 and 2 (compounds 8 and 9) isapproximately 1.5 pg based on S/N ratios.

Antibacterial Activity of F-Cip

Laboratory strain Pseudomonas aeruginosa (P. aeruginosa) ATCC 19660 isused to test the antimicrobial activity of modified ciprofloxacin. Thestrain is inoculated and grown at 37° C. for 18 h. The bacterialsuspension (300 mL) is added into 100 mL of nutrient broth and incubatedfor 2 h to reach a log growth phase. The resulting bacterial suspensionis adjusted to a concentration of ˜107 cfu/mL and 100 mL are placed intoeach well of a 96 well plate. Fifty microliters of F-Cip solution areadded into the 96 well plate and incubated at 37° C. for 18 h. Theinhibition of microbial growth is determined by measuring the absorbanceat 620 nm. The concentration that inhibited 50% of bacterial growth(IC50) is calculated based on absorbance readings from three independentexperiments.

Cytotoxicity of F-Cip and Modified Contact Lenses

Telomerase modified human corneal epithelial cells were seeded intowells of a 96 well plate with 100 mL KGM-2 medium (Lonza Ltd,Switzerland) and incubated at 37° C. until ˜80% confluence. Tenmicroliters of compounds 8-12 are added to the wells with finalconcentrations of 10, 5, 2.5, 1.25, 0.63, 0.31, 0.16, 0.08, 0.04, 0.02mM. Cells treated with 0.05% benzalkonium chloride (BAC) and without anytreatment served as controls. The plate was incubated at 37° C. for 18h, and cell viability was determined using a cell counting kit-8 assay.To determine the cytotoxicity of contact lenses, control and modifiedComfilcon A lenses are cut into small disks with a diameter of 6 mm andplaced individually into wells of the 96 well plate in duplicatefollowed by incubation at 37° C. for 18 h. All lenses are taken out, andcell viability was determined using a cell counting kit-8 assay (DojindoMolecular Technologies, Inc. Japan).

Ex Vivo Infection Model and Antimicrobial Activity of F-Cip LoadedContact Lenses.

Due to their similar size and anatomy with human eyes, porcine eyes havebeen adopted to test the effectiveness of contact lenses in ex vivomodels that had a mechanism to mimic the tear turnover. Fresh porcineeyes were purchased from a commercial slaughterhouse (Sioux-PremePacking Co., Chicago, Ill.). Upon arrival, eyelids and connectivetissues are removed and the porcine eyes are sanitized using 5%penicillin/streptomycin (Sigma-Aldrich, St. Louis, Mo.) for 1 h followedby washing thrice with PBS. Six scratch wounds with a 3×3 crosshatchpattern of length about 1 cm are created on each cornea using a 27Gneedle. The eyes are placed into holders and 100 mL of bacterialsolution containing 106 CFU (see “Antibacterial activity of F-Cip”section discussed above) applied to each cornea. The eyes are left atroom temperature in a biosafety cabinet for 5 h. After rinsing with 10mL of PBS, control and modified Comfilcon A contact lenses are placedindividually onto the eyes. Eyes without any lenses and eyes treatedwith 0.3% ciprofloxacin (Cip) solution every 15 min for 1 h served ascontrols. One eye without scratches but incubated with bacteria servedas an additional control. A custom-made apparatus is used to flowculture medium over the surface of the porcine eyes. This apparatusconsisted of a fluid reservoir containing DMEM (Life Technologies, Inc.Carlsbad, Calif.) and small diameter tubing positioned to create a flowrate of four 25 mL drops per min onto individual porcine eyes that areplaced directly underneath the tube outlet. The eyes were left for 12 h,and then rinsed with 10 mL PBS. The corneas were dissected and cut intopieces using a sterile scalpel followed by homogenizing using a LabGen125 homogenizer (Cole-Parmer, Vernon Hills, Ill.) for 1.5 min in 2 mLPBS. Five hundred microliters of the homogenate solution from eachsample was added into individual tubes with 6 mL nutrient broth, and themixtures were shaken at 250 rpm at 37° C. for 10 h. Then 100 mL of thebacterial suspension from each sample was taken out and added into a96-well plate in triplicate, and absorbance at 620 nm was measured andplotted as percentage of bacterial growth inhibition compared to thecorneas infected with bacteria but without any treatments.

Adsorption of Phosphatidylcholine and Cholesterol Oleate

The adsorption of lipids by modified contact lenses was determined usingradiolabelled lipids 3H-phosphatidylcholine and 14C-cholesterol oleatedetected by scintillation counting. Non-modified lenses are used ascontrol. Modified lenses are soaked in 3H-phosphatidylcholine (0.0005mg/mL) or 14C-cholesterol oleate (0.024 mg/mL) in phosphate bufferedsaline, or phosphate buffered saline alone as control. Lenses are placedindividually into glass vials containing the above lipids or phosphatebuffered saline and incubated for 8 h up to 24 h at 35° C. Each lens iswashed with phosphate buffered saline three times, and then counted in ascintillation counter. The experiment is repeated three times, and theamount of lipids adsorbed onto modified lenses is compared with theamount of lipids adsorbed onto non-modified lenses using student'st-test.

Adsorption of Lysozyme and Albumin

The adsorption of lysozyme and albumin onto modified lenses isdetermined using XPS. Modified lenses or non-modified lenses are soakedin lysozyme (1.9 mg/mL) or albumin (0.2 mg/mL) in phosphate bufferedsaline, or phosphate buffered saline alone as control. Lenses are placedindividually into glass vials containing the above proteins or phosphatebuffered saline and incubated for 8 h up to 24 h at 35° C. Each lens iswashed with PBS three times and vacuum-dried. The amount of proteinsadsorbed onto each lens is calculated by the nitrogen signal from XPS.The amount of proteins adsorbed onto modified lenses is compared withthe amount of protein adsorbed onto non-modified lenses using student'st-test.

Adsorption of Pseudomonas aeruginosa and Staphylococcus aureus

The number of bacteria adsorbed onto contact lenses was determined usingfluorescently stained Pseudomonas aeruginosa and Staphylococcus aureusdetected by fluorescence microscopy. Modified lenses were placedindividually into glass vials containing the above bacteria (107 cfu)and incubated for 8 h up to 24 h at 35° C. Non-modified lenses were usedas control. Each lens was washed with phosphate buffered saline threetimes, stained with hoechst, and observed using fluorescence microscopy.At least three imaging fields are randomly chosen for each lens sample.The number of bacteria adsorbed onto each lens per imaging field wascounted using ImageJ software. The experiments were repeated three timesand the amount of bacteria adsorbed onto modified lenses was comparedwith the amount of bacteria on non-modified lenses using student'st-test.

Example 5

Interaction of Fluorinated Zwitterionic Molecules with Different Typesof Lenses

To demonstrate the concept of using fluorous attractions to attachfluorinated molecules onto contact lenses with fluorine, a fluorinatedcarbon chain (FITC-C₈F₁₅) tagged with a fluorescent dye shown in FIG. 2is used as a model compound. The attachment of this compound can beeasily detected and quantified by its fluorescence signal. FITC is usedas control. Four types of contact lenses were tested and theirproperties are listed in Table 1. Among these 5 types of lens materials,Comfilcon A, Lotrafilcon B and Narafilcon A are silicone hydrogel,Ocufilcon D is conventional hydrogel, and Delefilcon A is a mixture ofboth materials. Notably, among these 4 types of lens materials, onlyComfilcon A has fluorine elements in its formulation.

TABLE 1 Properties of Lens Types Used Trade Name Biofinity Air OptixBiomedics 55 1-day Acuvue Dailies Total 1 UV TrueEye USAN Comfilcon ALotrafilcon B Ocufilcon D Narafilcon A Delefilcon A Manufacturer CooperAlcon Cooper Johnson & Alcon Vision Vision Johnson Water content % 48 3355 54 33 core, >80 surface Material Silicone Silicone Hydrogel SiliconeSilicon hydrogel hydrogel hydrogel hydrogel core non-silicone hydrogelsurface Atomic conc.  3.7  1.05  0  0 0 of surface FI %

Commercial lenses of the above 4 types were removed from blister packs,and blotted gently to remove excess solution. Contact lenses were placedindividually into glass vials and immersed in 1 mL of FITC-C₈F₁₅ or FITCsolution (15.6 μM in phosphate buffered saline) for 24 hours at roomtemperature. At the end of incubation, lenses were washed three timesusing phosphate buffered saline. The fluorescence of the incubationsolution as well as each wash solution was measured using a BMG LabTechFLUOstar Omega plate reader, and the amount of fluorescent moleculesattached onto the contact lenses is calculated as follows:M _(adsorbed onto contact lens) =M _(total) −M _(solution)where M_(total) is 15.6 nmole, and M^(solution) is the total amount offluorescent molecules in the incubation solution and wash solutions,which was calculated based on the fluorescence intensity for eachfluorescence dye.

As shown in FIG. 3 , Comfilcon A type lenses attached the greatestamount of FITC-C₈F₁₅ among the lenses tested. Furthermore, for theComfilcon A and Lotrafilcon B type lenses, the amount of FITC-C₈F₁₅ orFITC-C₈F₁₅ attached are significantly more than the amount of FITCattached (FITC-C₈F₁₅: Comfilcon A, 11 times and Lotrafilcon, B 6.5times; FITC-C₈F₁₅: Comfilcon A, 9 times and Lotrafilcon B, 5 times). Thestructural difference between FITC-C₈F_(n), and FITC is the fluorinatedcarbon chain (see FIG. 2 for their chemical structures). As only theComfilcon A and Lotrafilcon B type lens has fluorinated molecules in itsformulation, the above results indicated that the presence of fluorinein Comfilcon A and Lotrafilcon B type lens greatly facilitated theattachment of FITC-C₈F₁₅.

Example 6

Transparency of the Contact Lens After Drug Incorporation

FIG. 4A illustrates the transparency of contact lenses incubated withfluorinated ciprofloxacin compound F-Cip1 and control ciprofloxacin(Cip). Transmission of modified lenses remained remarkably close toabout 100% between 400 nm and 750 nm for bot Cip and F-Cip1 embeddedlenses. Similarly, as illustrated in FIG. 4B, lenses embedded with Cip,F-Cip1 and F-Cip2 show transmission profiles that significantly overlapwith that for a fresh unmodified lens between 200 nm and 800 nm. Asreported, ciprofloxacin (Cip) loaded lenses often exhibit precipitatesand lose their transparency. Therefore, the current simple method isadvantageous since it has addressed previous issues of loss oftransparency of Cip loaded lenses that often exhibited precipitates.

Example 7

The Release of Adsorbed Molecules from Contact Lenses Release of FITC-FCompounds from Lenses

Contact lenses with attached molecules prepared above were placed intoglass vials individually and immersed in 1 mL phosphate buffered salineat room temperature. On day 1, 2, 3, 8, and 15, the phosphate bufferedsaline solution was removed and the fluorescence of each solution wasmeasured. Then fresh phosphate buffered saline was added to each vial.The amount of FITC-C₈F₁₅ (compound 6) or FITC (compound 7) on each lenswas calculated as the initial amount on the lens minus the amountreleased into solution. FIGS. 5A-5D show release of FITC-C₈F₁₅ and FITCfrom Comfilcon A lens at 1 mL/h and 6 mL/h exchange rates. For solutionexchange rate of 1 mL/h, the release of unmodified FITC showed a typicalburst release profile, and reached plateau with ˜60% release within thefirst 2 h (FIG. 5B). The remaining 40% was likely on the lenses. Burstrelease is the major hurdle for clinical applications of current contactlens based drug delivery systems. Significantly, the release of FITC-Fcompound 6 showed a sustained profile with a nearly linear release forthe first 6 h (FIG. 5A). The release rate for compound 6 was determinedto be 10.5% per hour and reached ˜90% at 15 h (FIG. 5A). Upon increasingthe solution exchange rate to 6 mL/h, as expected, the release ofunmodified FITC quickly reached plateau within 50 min (FIG. 5D). On theother hand, release of FITC-F compound 6 showed a sustained linearrelease for 120 min (FIG. 5C). The release rate for compound 6 wasdetermined to be 26.5% per hour and reached plateau at 270 min (FIG.5C). Releasing the drug from contact lenses in such a sustained fashionis beneficial since it greatly improves drug adsorption efficiency andhence efficacy. Another advantage is that it will greatly reduce theamount of drug needed to achieve the same therapeutic efficacy, thusgreatly reducing toxicity.

These results showed the release rate increased with faster solutionexchange rate, which confirms that the release profiles from contactlens drug delivery model systems are dependent on the experimentalsetup, and care must be taken to compare results from studies withdifferent setups. The reported tear turnover rate (˜100 mL/h) in humansubjects varies significantly, and is much lower than the rates used inthis experiment. It should be pointed out that the solution in thisexperiment was changed hourly while tear turnover in the eye is acontinuous process, which could result in a different drug releaseprofile. Therefore, it is not feasible to predict the release profile ofthis system in vivo only based on the solution exchange rates.

Release of F-Cip Compounds from Lenses

FIG. 5E shows the release profile of F-Cip compounds 8 (F-Cip1) and 9(F-Cip2) and Cip (compound 12) from Comfilcon A lenses over a period of8 h with a solution exchange rate of 1 mL/h. Compound 12 exhibited atypical burst release profile, that is, most release occurred within thefirst hour and no more release after 3 h. The plateau was reached at˜60% release, and the remaining 40% were likely on the lenses.Significantly, both F-Cip compounds 8 and 9 exhibited a sustainedrelease profile that is very similar to the release profile of FITC-Fcompounds shown in FIGS. 5A to 5D. Although the percentage release ofF-Cip compounds 8 and 9 within the 8 h period was lower than that of Cip(Compound 12), the amount of F-Cip released was 6 times of that of Cipbecause the amount of F-Cip loaded onto lenses (˜68 nmol/lens) was muchmore than that of Cip (3.56±4.51 nmol/lens) as shown in FIG. 6 . Therelease profiles of F-Cip compounds 8 and 9 were very similar indicatingthat only one CF2 group difference in the structures of these twomolecules may not affect the overall fluorous interactions between F-Cipand contact lenses, which was also supported by the comparable amountsof F-Cip compounds 8 and 9 loaded onto Comfilcon A lenses. These resultsindicated that the fluorous attraction is so strong that it holds moremolecules for longer time than non-fluorous (Cip) interaction. This isadvantageous since greater drug adsorption efficiency results in higherdrug efficacy while at the same time greatly reducing the amount of drugneeded to achieve the same therapeutic effect, thereby minimizingtoxicity.

Example 8

Loading of Modified Ciprofloxacin into Comfilcon A Lenses

As Comfilcon A lenses present the greatest amount of fluorine in theformulation, this type of lens is used for the loading experiment ofF-Cip. As shown in FIG. 6 , the amount of compounds 8, 9, 10 and 11loaded onto lens is 67.96±11.50, 68.72±5.28, 108.36±5.95, and 83.31±8.33nmol/lens, which is about 19, 19, 30 and 23 times more than the amout ofCip loaded (3.56±4.51 nmol/lens) respectively. There are nostatistically significant differences for the loading of the compounds 8and 11 onto the contact lenses but all of them are significantly higherthan the amount of Cip.

This result demonstrates that the presence of even a short fluorous tagon the molecule greatly enhances its immobilization onto thefluorocarbon-containing contact lenses. The amount of compound 10 loadedon lenses was significantly more than that for compounds 8, 9 and 11,demonstrating that longer fluorocarbon chain (i.e. stronger interactionsbetween molecules and the lens) facilitated the loading onto lenses. Itis also remarkable that after modification, the lenses maintained theirtransparency.

As shown in FIGS. 4A and 4B, within the range of 200-800 nm,transmission of modified lenses remained ˜100% compared to fresh lenseswithout modification. Therefore, the current simple method has addressedthe previously reported issue of loss of transparency ofciprofloxacin-loaded lenses that often exhibited precipitates. Thestructure difference between compounds 8 and 9 and compound 12 is thefluorinated carbon chain, demonstrating that the presence of fluorine inthe molecule greatly facilitated its attachment to fluorine-containingcontact lens, which demonstrates that fluorine interactions play animportant role in the attachment of fluorinated compounds onfluorine-containing contact lenses. Importantly after modification, thelenses maintained their transparency (˜100%) within the visible lightrange (400-700 nm), compared to fresh lenses without modification.

TABLE 2 IC50 values of fluorinated ciprofloxacin against Pseudomonasaeruginosa (PA) ATCC 19660. No. m n IC50 (μM) F-Cip1, Compound 8 0 20.63 ± 0.15 F-Cip2, Compound 9 0 3 0.69 ± 0.62 F-Cip3, Compound 10 05 >40 F-Cip4, Compound 11 2 1 >40 Cip, Compound 12 — —  0.13 ± 0.035(n > 3)Antimicrobial Activity of Fluorinated Ciprofloxacin Loaded ContactLenses In Vitro Antimicrobial activity of F-Cip-Loaded Contact Lenses

The antimicrobial activity of fluorinated ciprofloxacin 1 (compound 8)and 2 (compound 9) loaded Comfilcon A lenses is tested against P.aeruginosa 19660. As shown in FIG. 7 , the lenses loaded with compounds8 and 9 exhibited 99.3% and 93.6% growth inhibition respectively, whichis significantly higher than 35.2% by Cip (compound 12) loaded lenses.Table 2 compares the IC50 values against P. aeruginosa 19660 for F-Cipcompounds with control Cip.

Although compounds 8 and 9 exhibited higher IC50 values (Table 2)against P. aeruginosa 19660 compared to ciprofloxacin, the amount ofF-Cip1 and F-Cip2 loaded onto Comfilcon A lenses is much more than Cip,which accounts for the need to add statistical analysis to be able tosay there is significantly more killing in F-Cip1 and F-Cip2 loadedlenses than Cip loaded lenses. Therefore, the antimicrobial activity ofdrug-loaded delivery systems does not entirely correlate with theantimicrobial efficacy of the loaded drug, but is also affected by theloaded amount.

It is expected that the antimicrobial activity of this drug deliverysystem is also dependent on the release profile of drug from vehicles.It is also worth noting that bacteria are also partially cleared fromthe eye with the continuous tear turnover, which may present a lesschallenging condition compared to the experimental condition tested herein which bacteria mare maintained in without other clearance mechanisms.Therefore, it is significant that this drug-loaded delivery systemexhibited nearly complete killing of bacteria in these demandingconditions, thus proving the antimicrobial efficacy of this drugdelivery system of the present invention.

Ex Vivo Infection Model and Antimicrobial Activity of F-Cip LoadedContact Lenses

Due to their similar size and anatomy with human eyes, porcine eyes havebeen adopted to test the effectiveness of contact lenses in ex vivomodels that had a mechanism to mimic the tear turnover. An ex vivoporcine eye infection model was developed, in which culture medium wasdripped onto each eye at a rate of four 25 μL drops/min to represent thetear turnover in vivo. Using this system, the antimicrobial activity ofcompound 8 and compound 12 loaded Comfilcon A lenses against P.aeruginosa 19660 was tested. Eyes that are infected with bacteria butnot exposed to contact lenses served as control.

As shown in FIG. 8 , for eyes with contact lenses modified with compound8 (F-Cip1), there was significant bacterial growth inhibition(86.8±13.9%) compared to the eyes with unmodified lenses (CL) andcompound 12 (Cip) modified lenses, which showed 29.7±12.7% and 37.5±8.2%bacterial growth inhibition, respectively. These results support datafrom the in vitro antimicrobial assay described above (illustrated inFIG. 7 ).

Porcine eyes infected with bacteria and treated with 0.3% Cip solutionshowed 98.1±4.3% bacterial growth inhibition, which is higher than theones wearing compound 8 modified lenses although this difference is notstatistically significant (p=0.70).

In clinical practice, a high concentration (0.3%) of Cip solution iscommonly used to treat eye infections and frequent administration isnecessary. For example, treatment of relatively mild bacterialconjunctivitis requires one to two drops four times a day, whiletreatment of sight threatening bacterial ulcers requires drops every 15min for the first 6 h. It should be noted that the amount ofciprofloxacin in the solution that is used for treatment is more than100 times the amount of compound 8 (67.96±11.50 nmol/lens, see FIG. 6 ),yet these two systems showed a comparable efficacy. Therefore, thisapproach could significantly reduce the amount of drug needed to treateye infections. For the eyes without scratch wounds, the number ofbacteria detected is much less than that for the control eyes indicatingthat bacteria did not penetrate the intact corneal epithelium and thosethat grew on top of the ocular surface are most likely washed awayduring the washing step. Overall, these results showed that contactlenses loaded with F-Cip exhibited antimicrobial activity in an ex vivomodel that represents tear clearance in vivo.

Cytotoxicity of Fluorinated Ciprofloxacin Loaded Contact Lenses

The cytotoxicity of compounds 8 and 9 loaded lenses was tested against ahuman corneal epithelial cell line. FIG. 9A shows cytotoxicity ofcompounds 8 and 9 and the unflorinated control compound 12 against humantelomerase corneal epithelial cells (hTCEpi) at a series ofconcentrations between 0 μM and 10 μM. Viability was about 100% with nosignificant difference among compound 8, compound 9 and compound 12indicating that modification of Cip with a fluorous tag did not changeits cytotoxicity. Benzalkonium chloride (BAC) known to be cytotoxickilled about 100% of cells and served as the positive control. FIG. 9Bshows that, cells incubated with untreated lenses exhibited decreasedviability compared to cells without any lenses (Ctrl), indicating thepotential damaging effect of overnight contact lens wear. For cellsincubated with lenses loaded with compounds 8 and 9, and controlcompound 12, there is no significant difference in toxicity compared tocells incubated with untreated lenses, indicating that loaded drug didnot exhibit a toxic effect on this cell line. The present invention iswell adapted to attain the ends and advantages mentioned as well asthose that are inherent therein. The particular embodiments disclosedabove are illustrative only, as the present invention may be modifiedand practiced in different but equivalent manners apparent to thoseskilled in the art having the benefit of the teachings herein.Furthermore, no limitations are intended to the details of constructionor design herein shown, other than as described in the claims below. Itis therefore evident that the particular illustrative embodimentsdisclosed above may be altered or modified and all such variations areconsidered within the scope and spirit of the present invention.

What is claimed is:
 1. A method for modifying a contact lens to attachdrugs to the contact lens, comprising the steps of: drying the contactlens, wherein the contact lens comprises fluorinated molecules, andwherein the contact lens has at least a 3.7% atomic concentration offluorinated molecules on surfaces of the contact lens; incubating thecontact lens with at least one fluorinated compound which is the drugfor attachment in a medium, wherein the at least one fluorinatedcompound attaches to the fluorinated molecules in the contact lensthrough a fluorous interaction to produce a modified contact lens, andwherein the at least one fluorinated compound is a fluorinatedophthalmic drug compound having the chemical structure:

wherein n is 0 or greater and m is 0 or greater; and rinsing themodified contact lens, wherein at least 68 nmol of the at least onefluorinated compound remain attached to the fluorinated molecules in themodified contact lens following rinsing.
 2. The method of claim 1,wherein the medium is a buffer or an organic solvent.
 3. The method ofclaim 2, wherein said buffer is phosphate buffered saline.
 4. The methodof claim 1, wherein the organic solvent is tetrahydrofuran, ethanol orisopropanol.
 5. The method of claim 1, wherein the step of incubatingthe contact lens is performed for about 4 hours to about 24 hours. 6.The method of claim 1, wherein the step of incubating the contact lensis performed at about room temperature.
 7. The method of claim 1,wherein said method prevents polymer chain rearrangement of the contactlens.
 8. The method of claim 1, wherein said method maintainstransparency of the contact lens.
 9. The method of claim 1, wherein saidcontact lens is resistant to microbial agents.